Fig. 5.

FoxO3a alters cyclin A expression. (A) Flow cytometry of lentiviral-infected KTC3 cells to examine silencing of FoxO3a, and transiently transfected FRO cells to examine overexpression of wt FoxO3a. Prior to analysis, cells were synchronized for 24 hours and then released for 72 hours. DNA content was analyzed using propidium iodide fluorescence and histograms were plotted of the percentage of cells in each phase of the cell cycle. (B) Western blot of the same cells indicate that when FoxO3a is silenced in KTC3 cells, cyclin A and B expression decreases, whereas overexpressing wt FoxO3a in FRO cells increases cyclin A and B expression. β-actin was used as a loading control. (C) QPCR confirms that only CCNA1 and A2 mRNA levels are decreased in FoxO3a-silenced KTC3 cells and only CCNA1 mRNA levels are increased in wt FoxO3a-overexpressing FRO cells. There is no change in CCNB1 and CCNB2 mRNA levels. Data are plotted as fold change ± s.d. compared with control, *P<0.05.